CTPsyn mutants.

<p>(A) Genetic map of CTPsyn and the insertion site of <i>p-element</i> in three mutants <i>CTPsyn^d6099</i>, <i>CTPsyn^e01207</i> and <i>CTPsyn^d07411</i>. (B) Comparison of the expressions of CTPsyn isoforms in larvae from <i>y w</i> and three <i>CTPsyn</i> mutants (<i>CTPsyn^d6099</i>, <i>CTPsyn^e01207</i> and <i>CTPsyn^d07411</i>). While dramatic decrease of isofrom C expression occurs in <i>CTPsyn^d6099</i> and <i>CTPsyn^e01207</i>, isoform A expression is hugely diminished in <i>CTPsyn^d07411</i>.</p

Expression profiles of <i>Drosophila</i> CTPsyn isoforms as revealed by qPCR.

<p>(A) embryos and larvae. (B) S2 cells and pupae. (C) Adult flies and tissues. WPP, white pre-pupae. P1–P4, pupal stages 1–4.</p

Overexpression of CTPsyn isoform C induces the assembly of cytoophidia in <i>Drosophila</i> follicle cells.

<p>(A, B) Cytoophidia revealed by CTPsyn-GFP in protein trap line CA06746. (C, D) Long cytoophidia are induced in stage-10 follicle cells overexpressing CTPsyn isoform C. (E, F) Each follicle cell contains only one long cytoophidium in a stage-7 egg chamber. The cell boundary is outlined by membrane protein Hu-li tai shao (Hts). (G) Within the same stage 10 egg chamber, cytoophidia in follicle cells overexpressing CTPsyn isoform C (GFP+ cells) are much longer and thicker than those in neighbouring wild-type follicle cells (GFP− cells). While the average length of cytoophidia in GFP− cells is only 3.97±0.61 µm (n = 41), cytoophidia in GFP+ cells are 20.76±5.60 µm long on average (n = 32). Note that cytoophidia in GFP+ cells in E are much longer than those in C and D, which could be due to different expression of GAL4 drivers in follicle cells (actin-GAL4 in C, D and tubulin-GAL4 in E). Scale bars, 20 µm.</p

CTPsyn mutants survive until larval stages.

<p><i>CTPsyn^d6099</i> homozygous mutants (−/− in A–E) and <i>CTPsyn^e01207</i> homozygous mutants (−/− in F–J) survive until 7 days after egg deposition (DAE) with slower growth compared to heterozygous mutants (+/−) or y w (+/+). (K–M) <i>CTPsyn^d07411</i> homozygous mutants (−/−) show severely delayed development, in comparison with heterozygous mutants (+/−and <i>y w</i> (+/+). <i>CTPsyn^d07411</i> (−/−) mutant larvae only survive until 5 days after egg deposition (DAE) with very little growth. Heterozygous mutants were balanced with TM6B Tb (+/− in all panels).</p

Clonal expression of CTPsyn domains in follicle cells using inducible driver.

<p>(A–F) Ectopic expression of UAS-SD-Venus disrupts endogenous cytoophidium formation in follicle cells (GFP+ cells), while cytoophidia are detectable in wild-type follicle cells (GFP− cells, within dotlined-area). (A–C) Surface view. (D–F) Lateral view. (G–I) Ectopic expression of UAS-Venus-GAT has no obvious effect on cytoophidium formation in follicle cells. Note that cytoophidia are detectable in both GFP+ (where the transgene is expressed) and GFP− cells (dotlined, wild-type). SD, synthetase domain; GAT, type 1 glutamine amidotransferase domain. Venus (C), cytoplasmic signal from Venus. GFP (N), nuclear signal from GFP. Scale bars, 20 µm.</p

The <i>Drosophila melanogaster</i> CG6854/CTPsyn gene locus encodes three isoforms.

<p>(A) Genome Browser view of the CG6854/CTPsyn gene locus (<a href="http://www.flybase.org" target="_blank">www.flybase.org</a>). In two protein trap lines (CA06746 and CA07332), GFP was trapped between the first and second exons of the CTPsyn isoform. (B) The protein map of three isoforms of CTPsyn. UTP binding sites are indicated by red triangles.</p

Gene ontology (GO) enrichment analysis of decidualization-related genes.

<p>(A) The union of human and mouse gene sets were analyzed using BiNGO software. Significantly enriched GOslim categories were highlighted with different colors representing different levels of significance. The size of each circle is correlated to the number of genes. (B) Comparative GO enrichment analysis for species-specific gene sets arranged in the biological process category (BP), the cellular component category (CC) and the molecular function category (MF), respectively. The analysis was applied to human and mouse gene sets, as well as 4 additional gene sets generated by set operations (union, intersection and difference) between them. The columns represent different gene sets, while the rows represent statistically significant GO terms.</p

Ectopic expression of transgenes in egg chambers using <i>MTD-GAL4</i> driver.

<p>(A–C) UAS-Venus-N-term which consists of the 56 amino acids shows dispersed cytoplasmic expression. (D–I) UAS-Trun isoC-Venus forms punctate structures in the cytoplasm. The punctate structures are more visible in egg chambers at later stages (G–I) than in those at early stages (D–F). (J–L) UAS-Venus-Trun isoC shows dispersed distribution in the cytoplasm. (M–O) Ectopic expression of UAS-SD-Venus disrupts endogenous cytoophidium formation in the germline cells. Note that there is no expression of UAS-SD-Venus in follicle cells in which cytoophida are not affected. (P–R) UAS-GAT-Venus shows disperse distribution in the cytoplasm. See <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003256#pgen.1003256.s006" target="_blank">Figure S6</a> for the structures of the constructs. N-term, a N-terminal segment (56-aa) of CTPsyn isoform C; Trun IsoC, truncated isoform C. SD, synthetase domain; GAT, type 1 glutamine amidotransferase domain. Scale bars, 20 µm.</p

Pathway enrichment analysis of decidualization-related genes.

<p>(A) The figure shows the significantly enriched pathways identified by using DAVID online tools. The bars represent the enrichment p-value at logarithmic scale. (B) Visualization of the LIF-STAT signaling pathway. Nodes represent genes. The node color indicates the status of the gene as specific to human (red), specific to mouse (green), or shared by both (blue). Edges represent gene dependences derived from KEGG pathway database. Genes without a direct interaction with others are not included. This graph is generated using the Cytoscape software.</p

Systematic Analysis of the Molecular Mechanism Underlying Decidualization Using a Text Mining Approach

<div><p>Decidualization is a crucial process for successful embryo implantation and pregnancy in humans. Defects in decidualization during early pregnancy are associated with several pregnancy complications, such as pre-eclampsia, intrauterine growth restriction and recurrent pregnancy loss. However, the mechanism underlying decidualization remains poorly understood. In the present study, we performed a systematic analysis of decidualization-related genes using text mining. We identified 286 genes for humans and 287 genes for mice respectively, with an overlap of 111 genes shared by both species. Through enrichment test, we demonstrated that although divergence was observed, the majority of enriched gene ontology terms and pathways were shared by both species, suggesting that functional categories were more conserved than individual genes. We further constructed a decidualization-related protein-protein interaction network consisted of 344 nodes connected via 1,541 edges. We prioritized genes in this network and identified 12 genes that may be key regulators of decidualization. These findings would provide some clues for further research on the mechanism underlying decidualization.</p></div